Compounds With Matrix-Metalloproteinase Inhibitory Activity and Imaging Agents Thereof

ABSTRACT

Novel compounds and pharmaceutical compositions having MMP inhibitory activity are disclosed, which have been found to be particularly useful in the prevention, treatment and diagnostic imaging of diseases associated with an unpaired activity of MMP, amongst others MMP-2, MMP-8, MMP-9 and/or MMP-13 to name a few. The compounds of the present invention are useful for the prevention, the treatment and the in vivo diagnostic imaging of a range of disease states (inflammatory, malignant and degenerative diseases) where specific matrix metalloproteinases are known to be involved.

RELATED APPLICATIONS

This application claims priority to provisional patent application U.S.Ser. No. 61/616,494; filed on Mar. 28, 2012; the entire contents ofwhich are incorporated herein by reference. This application is acontinuation-in-part application of, and claims priority to, U.S. Ser.No. 13/155,558; filed on Jun. 8, 2011; the entire contents of which areincorporated herein by reference. This application is acontinuation-in-part application of, and claims priority to,PCT/EP2011/062211; filed on Jul. 18, 2011; the entire contents of whichare incorporated herein by reference.

FIELD OF THE INVENTION

The present invention relates to the field of therapeutic, diagnosticand imaging agents and more specifically to compounds that areinhibitors of matrix-metalloproteinases (MMPs).

BACKGROUND

The matrix-metalloproteinases (MMPs) are a family of at least 20zinc-dependent endo-peptidases which mediate degradation, or remodelingof the extracellular matrix (ECM). Together, the members of the MMPfamily can degrade e.g. components of the blood vessel wall and play amajor role in both physiological and pathological events that involvethe degradation of components of the ECM. Since the MMPs can interferewith the cell-matrix interactions that control cell behavior, theiractivity affects processes as diverse as cellular differentiation,migration, proliferation and apoptosis. The negative regulatory controlsthat finely regulate MMP activity in physiological situations do notalways function as they should. Inappropriate expression of MMP activityis thought to constitute part of the pathological mechanism in severaldisease states. MMPs are therefore targets for therapeutic inhibitors inmany inflammatory, malignant and degenerative diseases. Consequently, itis believed that synthetic inhibitors of MMPs may be useful in thetreatment of many inflammatory, malignant and degenerative diseases.Furthermore, it has been suggested that inhibitors of MMPs may be usefulin the diagnosis of these diseases.

The compounds of the present invention are useful for the prevention,the treatment and the in vivo diagnostic imaging of a range of diseasestates (inflammatory, malignant and degenerative diseases) wherespecific matrix metalloproteinases are known to be involved. Theseinclude: (a) atherosclerosis; (b) CHF; (c) cancer; (d) arthritis; (e)amyotrophic lateral sclerosis; (f) brain metastases; (g) cerebrovasculardiseases; (h) Alzheimer's disease; (i) neuroinflammatory diseases; (j)COPD; (k) eye pathology; (l) skin diseases.

SUMMARY

Novel compounds having MMP inhibitory activity are disclosed, which havebeen found to be particularly useful in the prevention, treatment anddiagnostic imaging of diseases associated with an unpaired activity ofMMP, amongst others MMP-2, MMP-8, MMP-9 and/or MMP-13 to name a few.

Another aspect of the present invention relates to pharmaceuticalcompositions useful in prevention, treatment and diagnostic imaging ofdiseases associated with an unpaired activity of MMP.

The compounds of the present invention are useful for the prevention,the treatment and the in vivo diagnostic imaging of a range of diseasestates (inflammatory, malignant and degenerative diseases) wherespecific matrix metalloproteinases are known to be involved.

The compounds of the present invention may be “cold” or comprise “hot”radioactive atoms or molecules. Imaging compounds disclosed herein mayhave radiolabels selected from the group consisting of ¹¹C, ¹³N, ¹⁵O,¹⁸F, ⁶¹Cu, ⁶²Cu, ⁶⁴Cu, ⁶⁷Cu, ⁶⁸Ga, ¹²⁴I, ¹²⁵I, ¹³¹I, ⁹⁹Tc, ⁷⁵Br, ¹⁵³Gdand ³²P.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1A is a graph showing MMP activity measurement in mouse tumor,muscle, brain and aorta.

FIG. 1B is a graph showing correlation with fluorescence intensity afterM005 injection, 4 h uptake.

FIG. 2 is a graph showing correlation with fluorescence intensity afterM005 injection, 4 h uptake (all experiments)

FIGS. 3-5 are mouse scans before and 4 hours after M005 injection.

FIG. 6 shows biodistribution of M005: tumor and tissue scans (4 h uptaketime, Cy5 channel, with and without tracer injection).

FIG. 7 shows tissue sections (10 μm) scan after M005 injection (uptaketime: 4 h (Cy5.5 channel, Alpha-Innotech).

FIG. 8 shows M005 staining at different concentrations on tumorsections.

FIG. 9 shows average MMP activity measurement in mouse tissues.

DETAILED DESCRIPTION

The compounds of the invention may be used as therapeutics and/or invivo diagnostic agents. In a further aspect, the present compounds maybe used to prevent and/or treat pathological conditions associated withunpaired expression of matrix-metalloproteases in human and animal, inparticular mammals.

In another aspect, the present invention relates to compounds that arelabeled with a radionuclide such as an ¹⁸F atom for use as a diagnosticor imaging agent, in particular as an in vivo diagnostic or imagingagent and more in particular as diagnostic or imaging agent for PositronEmission Tomography (PET) or SPECT.

The compounds may be used to visualize, assess and quantify MMP-activityin cells and tissues. Preferably, molecular imaging agents are forvisualizing and quantifying MMP-activity in mammalian cells and tissues,including human cells and tissues. This may comprise analyzing cellularor tissue radioactivity content, or the rate of uptake, displacement,dissociation or partitioning.

The compounds of the invention may be used in the prevention ortreatment of pathological conditions associated with a dysregulatedexpression of matrix metalloproteinase in human and animal.

The radiolabeled compounds of the invention may be used for diagnosingpathological conditions associated with a dysregulated expression ofmatrix metalloproteinase in human and animal.

The pathological condition may be selected from the group consisting ofcardiovascular diseases, inflammatory diseases, autoimmune diseases andmalignant diseases. In particular the cardiovascular diseases may beselected from atherosclerosis and congestive heart failure. Inflammatorydisease is chronic obstructive pulmonary disease. Autoimmune diseasesare diabetes mellitus type 1, rheumatoid arthritis, multiple sclerosis,and malignant diseases are cancers.

The diagnostic imaging compound of the invention permits theidentification of active plaque burden, which allows risk stratificationof patients with known or suspected coronary artery disease, i.e.patients with pain or a history of pain, or identified as high risk butasymptomatic.

In addition, the diagnostic imaging agents of the invention permitidentification of vulnerable plaques in symptomatic patients, which mayallow identification of high risk of acute myocardial infarction orstroke irrespective of stenosis and permits immediate riskstratification when the patient suffers from chest pain Furthermore,angioplasty of vulnerable plaques is high risk, and may lead to embolismof the artery tree post surgery. Thus imaging of this subtype of plaquesmay help reduce post-surgical complication.

Suitable preparations include for example tablets, capsules,suppositories, solutions, —particularly solutions for injection (s.c,i.v., i.m.) and infusion—syrups, elixirs, solution for inhalation.

The invention relates to pharmaceutical compositions comprising aneffective amount, especially an amount effective in the treatment of oneof the above-mentioned disorders, of the active ingredient together withpharmaceutically acceptable carriers that are suitable for topical,enteral, for example oral or rectal, or intravenous or parenteraladministration and that may be inorganic or organic, solid or liquid.They are used for oral administration especially tablets or gelatincapsules that comprise the active ingredient together with diluents, forexample lactose, dextrose, sucrose, mannitol, sorbitol, cellulose and/orglycerol, and/or lubricants, for example silica, talc, stearic acid orsalts thereof, such as magnesium or calcium stearate and/or polyethyleneglycol. Tablets may also comprise binders, for example magnesiumaluminum silicate, starches, such as corn, wheat or rice starch,gelatin, methylcellulose, sodium carboxymethylcellulose and/orpolyvinylpyrrolidone, and, if desired, disintegrators, for examplestarches, agar, alginic acid or a salt thereof, such as sodium alginateand/or effervescent mixtures or adsorbents, dyes, flavorings andsweeteners.

It is also possible to use the pharmacologically active compounds of thepresent invention in the form of intravenously and parentallyadministrable compositions or in the form of infusion solutions. Suchsolutions are preferably isotonic aqueous solutions or suspensionswhich, for example in the case of lyophilized compositions that comprisethe active ingredient alone or together with a carrier, for examplemannitol, can be made up prior to use. The pharmaceutical compositionsmay be sterilized and/or may comprise excipients, for examplepreservatives, stabilizers, wetting agents and/or emulsifiers,solubilizes, salts for regulating the osmotic pressure and/or buffers.The present pharmaceutical compositions are prepared in a manner knownper se, for example by means of conventional mixing, granulating,confectioning, dissolving or lyophilizing processes, and compriseapproximately from 1% to 95%, especially from approximately 1% toapproximately 20%, active ingredient (s).

In one embodiment, the invention is a compound of the formula:

and pharmaceutically acceptable salts and stereoisomers thereof,wherein:

R₁ is selected from the group consisting of: C₁₋₆alkyl, C₁₋₆alkyl-halo,protecting group or leaving group;

A is a bond or is (CH₂)₁₋₅;

R₃ is selected from the group consisting of: OH, NH₂, protecting groupor leaving group;

R₄ is a bond or is at least one selected from the group consisting of:aryl, heteroaryl,

-   -   C₁₋₂₀alkyl, wherein at least one C of C₁₋₂₀alkyl is optionally        replaced by O, S, NH, C(O), aryl or heteroaryl and wherein at        least one H of C₁₋₂₀alkyl is optionally replaced with OH, NH₂,        COOH,        -   wherein at least one C of C₁₋₈alkyl is optionally replaced            by O, S, C(O), aryl, heteroaryl or NH,

R₅ is selected from the group consisting of: H, halo, CH₃, NH₂, OH, SO₂,NO₂, —O—C₁₋₆alkyl and

wherein at least one H or halo is optionally replaced with aradionuclide.

In one embodiment, A is a bond.

In one embodiment, R₁ is C₁₋₆alkyl.

In one embodiment, R₃ is OH.

In one embodiment, R₅ is a halo or a radionuclide.

In one embodiment, R₄ is C₁₋₂₀alkyl, wherein at least one C ofC₁₋₂₀alkyl is optionally replaced by O, S, NH, C(O).

In one embodiment, R₄ is aryl.

In one embodiment, R₄ is:

In one embodiment, R₄ is C₁₋₂₀alkyl, wherein: at least one C ofC₁₋₂₀alkyl is replaced by O, at least one C of C₁₋₂₀alkyl is replaced byC(O)NH, at least one C of C₁₋₂₀alkyl is replaced by aryl, at least one Cof C₁₋₂₀alkyl is replaced by heteroaryl, wherein at least one H of thearyl is replaced by COOH.

In one embodiment, R₄ is —[(CH₂)₂—O]₁₋₆—.

In one embodiment, R₅ is:

In one embodiment, the heteroaryl is a triazole.

In one embodiment, at least one H of C₁₋₂₀alkyl is replaced with

wherein Z is a halo or a radionuclide.

In one embodiment, R₄ is

In one embodiment, the invention is a compound of:

wherein Z is selected from the group consisting of: NH₂, OMe, halo orradionuclide.

In one embodiment, the invention is a compound of:

In one embodiment, the invention is a compound of:

In one embodiment, the invention is a compound of:

wherein Z is a halo or a radionuclide.

In one embodiment, the invention is a compound of:

wherein Z is a halo or a radionuclide.

In one embodiment, the invention is a compound that is:

and pharmaceutically acceptable salts and stereoisomers thereof,wherein:

each R₁ is independently selected from the group consisting of:C₁₋₆alkyl, C₁₋₆alkyl-halo, protecting group or leaving group;

each A is independently a bond or is (CH₂)₁₋₅;

each R₃ is independently selected from the group consisting of: OH, NH₂,protecting group or leaving group;

each R₆ is independently a bond or is at least one selected from thegroup consisting of: aryl, heteroaryl, or

-   -   C₁₋₂₀alkyl, wherein at least one C of C₁₋₂₀alkyl is optionally        replaced by O, S, NH, C(O), aryl or heteroaryl and wherein at        least one H of C₁₋₂₀alkyl is optionally replaced with OH, NH₂,        C₁₋₈alkyl, C₁₋₈alkyl-halo,        -   wherein at least one C of C₁₋₈alkyl is optionally replaced            with at least one of O, S, C(O), NH, aryl and heteroaryl;

each R₇ is independently a bond or is at least one selected from thegroup consisting of: CH₂, NH, C(O)NH, C₁₋₆alkyl, wherein at least one Cof C₁₋₆alkyl is optionally replaced by O, S, NH, C(O) and wherein atleast one H of C₁₋₆alkyl is optionally replaced by OH, NH₂, SO₂,C₁₋₈alkyl, C₁₋₈alkyl-halo,

wherein at least one C of C₁₋₈alkyl is optionally replaced by O, S,C(O), aryl, heteroaryl or NH,

R₈ is a bond or is at least one selected from the group consisting of:O, NH, S, aryl, heteroaryl, wherein at least one H of aryl or heteroarylis optionally replaced with C₁₋₈alkyl, C₁₋₈alkyl-halo,

wherein at least one C of C₁₋₈alkyl is optionally replaced with at leastone of O, S, C(O), NH, aryl and heteroaryl;

(CH₂)₁₋₆, wherein at least one C of (CH₂)₁₋₆ is optionally replaced byO, NH or S;

wherein at least one H or halo is optionally replaced with aradionuclide.

In one embodiment, each A is a bond.

In one embodiment, each R₃ is OH.

In one embodiment, each R₁ is CH₃.

In one embodiment, each R₆ is a bond.

In one embodiment, R₈ is O.

In one embodiment, each R₇ is C₁₋₆alkyl.

In one embodiment, each R₆ is C₁₋₂₀alkyl, wherein at least one C ofC₁₋₂₀alkyl is optionally replaced by O, S, NH, C(O), aryl or heteroaryland wherein at least one H of C₁₋₂₀alkyl is optionally replaced with OH,NH₂, C₁₋₈alkyl, C₁₋₈alkyl-halo, wherein at least one C of C₁₋₈alkyl isoptionally replaced with at least one of O, S, C(O), NH, aryl andheteroaryl.

In one embodiment, each R₆ is —[(CH₂)₂—O]₁₋₆—.

In one embodiment, each R₆ is:

In one embodiment, each R₇ is C₁₋₆alkyl, wherein at least one C ofC₁₋₆alkyl is optionally replaced by O, S, NH, C(O) and at least one H isreplaced with:

wherein Z is a halo or a radionuclide.

In one embodiment, each R₇ is CH₂C(O)NH.

In one embodiment, R₈ is aryl.

In one embodiment, at least one H of aryl is replaced withC₁₋₈alkyl-halo, wherein at least one C of C₁₋₈alkyl-halo is optionallyreplaced with heteroaryl.

In one embodiment, the invention is a compound that is:

and pharmaceutically acceptable salts and stereoisomers thereof.

In one embodiment, the invention is compound that is:

and pharmaceutically acceptable salts and stereoisomers thereof,

wherein Z is a halo or a radionuclide.

In one embodiment, the invention is compound that is:

and pharmaceutically acceptable salts and stereoisomers thereof,

wherein Z is a halo or a radionuclide.

In one embodiment, the invention is compound that is:

and pharmaceutically acceptable salts and stereoisomers thereof,wherein:

R₁ is selected from the group consisting of: C₁₋₆alkyl, C₁₋₆alkyl-halo,protecting group or leaving group;

A is a bond or is (CH₂)₁₋₅;

R₃ is selected from the group consisting of: OH, NH₂, protecting groupor leaving group;

R₁₀ is selected from the group consisting of: an alkyne, an azide,C₁-C₆-alkyne, C₁-C₆-azide, wherein at least one C is optionally replacedby NH, C(O), S or O.

In one embodiment, the invention is compound that is:

and pharmaceutically acceptable salts and stereoisomers thereof,wherein:

R₁ is selected from the group consisting of: C₁₋₆alkyl, C₁₋₆alkyl-halo,protecting group or leaving group;

A is a bond or is (CH₂)₁₋₅;

R₃ is selected from the group consisting of: OH, NH₂, protecting groupor leaving group;

R₉ is a bond or is at least one selected from the group consisting of:aryl, heteroaryl,

-   -   C₁₋₂₀alkyl, wherein at least one C of C₁₋₂₀alkyl is optionally        replaced by O, S, NH, C(O), aryl or heteroaryl and wherein at        least one H of C₁₋₂₀alkyl is optionally replaced with OH, NH₂,        COOH, C₁₋₈alkyl, C₁₋₈alkyl-halo,        -   wherein at least one C of C₁₋₈alkyl is optionally replaced            by O, S, C(O), aryl, heteroaryl or NH,

R₁₀ is selected from the group consisting of: an alkyne, an azide,C₁-C₆-alkyne, C₁-C₆-azide, wherein at least one C is optionally replacedby NH, C(O), S or O.

In one embodiment, the invention is compound that is:

and pharmaceutically acceptable salts and stereoisomers thereof,

wherein:

R₁ is selected from the group consisting of: C₁₋₆alkyl, C₁₋₆alkyl-halo,protecting group or leaving group;

A is a bond or is (CH₂)₁₋₅;

R₃ is selected from the group consisting of: OH, NH₂, protecting groupor leaving group;

R₉ is a bond or is at least one selected from the group consisting of:aryl, heteroaryl,

-   -   C₁₋₂₀alkyl, wherein at least one C of C₁₋₂₀alkyl is optionally        replaced by O, S, NH, C(O), aryl or heteroaryl and wherein one H        of C₁₋₂₀alkyl is optionally replaced with OH, NH₂, COOH,        C₁₋₈alkyl, C₁₋₈alkyl-halo,        -   wherein at least one C of C₁₋₈alkyl is optionally replaced            by O, S, C(O), aryl, heteroaryl or NH,

R₁₀ is selected from the group consisting of: an alkyne, an azide,C₁-C₆-alkyne, C₁-C₆-azide, wherein at least one C of C₁-C₆ is optionallyreplaced by NH, C(O), S or O;

-   -   R₁₁ is selected from the group consisting of: H, CH₃, NH₂, OH,        SO₂, NO₂, —O—C₁₋₆alkyl and

In one embodiment, the invention is compound that is:

and pharmaceutically acceptable salts and stereoisomers thereof,wherein:

each R₁ is independently selected from the group consisting of:C₁₋₆alkyl, C₁₋₆alkyl-halo, protecting group or leaving group;

each A is independently a bond or is (CH₂)₁₋₅;

each R₃ is independently selected from the group consisting of: OH, NH₂,protecting group or leaving group;

each R₆ is independently a bond or is at least one selected from thegroup consisting of: aryl, heteroaryl, or

-   -   C₁₋₂₀alkyl, wherein at least one C of C₁₋₂₀alkyl is optionally        replaced by O, S, NH, C(O), aryl or heteroaryl and wherein at        least one H of C₁₋₂₀alkyl is optionally replaced with OH, NH₂,        C₁₋₈alkyl, C₁₋₈alkyl-halo,        -   wherein at least one C of C₁₋₈alkyl is optionally replaced            with at least one of O, S, C(O), NH, aryl and heteroaryl;

each R₁₂ is independently a bond or is C₁₋₆alkyl, wherein at least one Cof C₁₋₆alkyl is optionally replaced by O, S, NH, C(O) and wherein atleast one H of C₁₋₆alkyl is optionally replaced by OH, NH₂, SO₂,C₁₋₈alkyl, C₁₋₈alkyl-halo, azide or alkyne,

wherein at least one C of C₁₋₈alkyl is optionally replaced by O, S,C(O), aryl, heteroaryl or NH,

R₈ is a bond or is at least one selected from the group consisting of:O, NH, S, aryl, heteroaryl, wherein at least one H of aryl or heteroarylis optionally replaced with C₁₋₈alkyl, C₁₋₈alkyl-halo,

wherein at least one C of C₁₋₈alkyl is optionally replaced with at leastone of O, S, C(O), NH, aryl and heteroaryl;

(CH₂)₁₋₆, wherein at least one C of (CH₂)₁₋₆ is optionally replaced byO, NH or S.

In a further aspect the present invention therefore provides a methodfor the molecular imaging of MMP-activity which comprises the steps of:

-   -   a. contacting said cells or tissues with a radiolabeled compound        of the present invention or composition of the present invention        and    -   b. detecting said MMP-activity.

Preferably, the step of detecting said MMP-activity comprises the stepsof positioning the subject within the detection field of a detectiondevice and detecting said compounds in the subject with said detectiondevice.

This method may be carried out also in vitro by contacting the cells ortissues with a compound or composition of the present invention byexposing, incubating, touching, associating or making the compoundaccessible to the cells or tissue. When the compound or composition ofthe present invention is radiolabeled, said MMP-activity can be detectedin vitro, ex vivo and in vivo using any appropriate radiation detectiondevice.

The compounds or compositions may be administered to a subject by anysuitable administration method (oral, injection (intravenous (IV),intramuscular (IM), and subcutaneous, parenteral), via inhalation,etc.). Preferably, the compounds are administered intravenously.

When the compound or composition of the present invention isradiolabeled, said MMP-activity may be detected using a radiationdetection device. Said radiation detection device may include a PositronEmission Tomography (PET) scanner or a Single Photon Emission ComputedTomography (SPECT) scanner. Preferably, said radiation detection deviceis a Positron Emission Tomography (PET) scanner combined with ComputerTomography (PET/CT) or Magnetic Resonance Tomography (PET/MR). Said PETscanner can detect pairs of gamma rays, emitted indirectly bypositron-emitting radioisotopes such as ¹⁸F to produce a reconstructed3D image of the radioactivity distribution within tissues. PET cantherefore be used to produce a 3D image of the distribution of theradiolabeled compounds and compositions of the present invention withinmammalian or human tissues.

TABLE 1 shows embodiments of MMP inhibitor/imaging compounds: MMP-2MMP-8 IC50, MMP-9 IC50, MMP-13 IC50, ID Structure MW cLogP IC50, nM nMnM nM M001

504 3.2  92% 100 nM  99% 100 nM M002

492 3.4 M003

1458  5.2  97% 100 nM  97% 100 nM M004

1237 25  83% 100 nM  91% 100 nM M005

1756  2  97% (100 nM)  2.1 100% 100 nM Cy5.5

917  0% (1 uM)  0% (0.1 uM)  1% (1 uM)  0% (0.1 uM)  17% (1 uM)  12%(0.1 uM)  7% at 1 uM,  0% at 0.1 uM M006

475 M007

837 64% (10 nM) 95(100 nM)  94% (10 nM) 100% (100 nM)  89% (10 nM) 100%(100 nM)  96% (10 nM)  99% (100 nM) M008

583 82% (10 nM)  95% (10 nM)  78% (10 nM)  82% (10 nM) M009

1398 59% (10 nM)  46% (10 nM)  48%  64% (10 nM) M010

1454 28% (10 nM)  13% (10 nM)  2% (10 nM)  10% (10 nM) M011

1198 50% (10 nM)  57% (10 nM)  55% (10 nM)  75% (10 nM) M012

 1% (10 nM)  0% (10 nM)  0% (10 nM)  0% (10 nM) M013

2 66% (10 nM)  84% (10 nM)  67% (10 nM)  79% (10 nM) M014

455.6 2.3 M015

711.8 2 M016

449.5 1.67 M017

1245 M018

1481 M019

1644 M020

1033 M021

792.9 0.77 M022

1739 0.77 M023

881 M024

1422Any of the foregoing compounds may include a fluorophore or radiolabel.For example, M005 may be:

Embodiments of the present invention are shown below:

As one example, the M007 compound shows good MMP inhibition:

One example of the preparation of M007 analogs is shown below:

One example of a compound of the present invention is the Cy5.5-[F18]-MMP tracer M005:

FIG. 6 shows biodistribution of M005: tumor and tissue scans (4 h uptaketime, Cy5 channel, with and without tracer injection). M005 shows uptakeabove background fluorescence of tissue. The brain was used asbackground tissue since there is no uptake of the tracer into the brain.

MMP Activity Measurement:

The activities were determined using Fluorimetric SensoLyte® 390 GenericMMP Activity Kit from Anaspec (cat #72202). This assay kit can detectthe activity of several MMPs such as MMP-1, 2, 7, 8, 9, 13, 14, 15, 16and 24.

For screening MMP compounds, activated 2 nM of recombinant MMP-2, 8, 9or 13 are pre-incubated with MMP compound (inhibitor) for 30 min at 37°C. The substrate is added and the inhibition of MMP activity isexpressed as % of total activity (without the inhibitor incubation).

For measuring the MMPs activity in biological samples, the substrate ismixed with sample lysate and immediately start measuring fluorescenceintensity at Ex/Em=330 nm/390 nm continuously. The result is normalizedby protein concentration.

The following describes an example of how at least one of the compoundsof the present invention may be prepared. Other methods may be used andthe below methods may be used to synthesize other compounds describedherein.

Synthesis of M005 precursor

To a vial charged with compound 1 (S)-6-amino-2-(2-azidoacetamido) N (2(2 (2(2-(4-((N—((R)-1-(hydroxyamino)-3-methyl-1-oxobutan-2-yl)-4-methoxyphenylsulfonamido)methyl)-1H-1,2,3-triazol-1-yl)ethoxy)ethoxy)ethoxy)ethyl)hexanamide(5.32 mg, 6.14 umol) and Cy5.5-NHS ester (6.60 mg, 5.85 umol) in DMF(0.2 ml), was added triethylamine DMF solution (0.1 M, 120 ul, 12 umol)via a micro syringe drop wise. The mixture was stirred at roomtemperature in dark for 30 min until LCMS indicated complete consumptionof the Cy5.5-NHS ester. The reaction was solution was diluted with HCl(aq 0.1 M, 0.4 mL), injected to a semi preparative reversed-phase HPLCwith Phenomenex Gemini C-18 (250×21.20 mm). The fraction containing M005precursor was eluted with mobile phase (0.15% TFA in acetonitrile andwater, 30 min gradient method 10% to 90% acetonitrile in water) andafforded M005 precursor (4.80 mg, 2.88 umol, yield 49.2%) afterlyophilization. ¹H NMR (400 MHz, D₂O): δ 8.34-8.32 (d, J=9.2 Hz, 2H),8.23 (s, 2H), 7.89 (s, 2H), 7.68-7.52 (m, 2H), 7.24-7.10 (m, 3H), 6.89(d, J=8.8 Hz, 2H), 6.26 (d, J=8.8 Hz, 2H), 5.98 (m, 1H), 5.70-5.48 (m,2H), 4.01 (d, J=12.0 Hz, 1H), 3.90 (m, 2H), 3.65-3.55 (m, 2H), 3.46 (s,4H), 3.22-3.16 (m, 5H), 3.07 (d, J=10.8 Hz, 1H), 2.93-2.91 (m, 10H),2.82-2.70 (m, 2H), 2.40-2.35 (m, 2H), 1.65-1.52 (m, 3H), 1.42-1.35 (m,12H), 1.20 (m, 2H), 1.00 (m, 4H), 0.88-0.55 (m, 10H), 0.23 (d, J=6.0 Hz,3H), 0.08 (d, J=6.4 Hz, 3H). Mass Spec (lo-res): Calc'd forC₇₂H₉₃N₁₃O₂₃S₅: 1667.5. Found: 835.0 (M+2H⁺)/2.

Synthesis of M005 Reference Standard

Compound 1 (5.00 mg, 6 umol) was added to a vial contains CuSO₄ (aq 0.1M, 0.1 ml, 10 umol), sodium ascorbate (0.50 mg, 1.7 umol) in DMF (0.2mL). 5-fluoropent-1-yne (8.60 mg, 0.1 mmol) in THF (20 uL) was added.The mixture was stirred at room temperature for 30 min until LCMSindicated the full conversion of the reaction. The mixture was purifiedon HPLC with the same condition described above. The fraction containsclick reaction product (4.0 mg) was dried on lyophilizer overnight andused directly. To a vial contains the click product (1.69 mg, 1.97umol), was added Cy5.5-NHS ester (2.00 mg, 1.97 umol) in DMF (0.2 ml)and triethylamine (0.1M in DMF, 20 ul). The reaction was stirred in darkfor 48 hours, diluted with HCl (aq 0.1 M, 0.3 ml), and purified on HPLCusing the same condition described above. The fraction contains M005reference standard was dried by overnight lyophilization to afford M005reference standard (2.60 mg, 1.48 umol, yield 75%) ¹H NMR (400 MHz, D₂O)δ 8.34-8.32 (d, J=9.2 Hz, 2H), 8.23 (s, 2H), 7.89 (s, 2H), 7.68-7.52 (m,2H), 7.24-7.10 (m, 4H), 6.89 (d, J=8.8 Hz, 2H), 6.26 (d, J=8.8 Hz, 2H),5.98 (m, 1H), 5.70-5.48 (m, 2H), 4.70 (s, 2H), 4.01-3.90 (m, 5H),3.65-3.46 (m, 4H), 3.22-3.16 (m, 5H), 3.07 (d, J=10.8 Hz, 1H), 2.93-2.91(m, 10H), 2.82-2.70 (m, 2H), 2.40-2.35 (m, 2H), 2.20-2.18 (m, 2H),1.65-1.52 (m, 3H), 1.42-1.35 (m, 12H), 1.20 (m, 2H), 1.00 (m, 4H),0.88-0.55 (m, 10H), 0.23 (d, J=6.0 Hz, 3H), 0.08 (d, J=6.4 Hz, 3H). ¹⁹FNMR (376 MHz, D₂O): −219.07 (tt, J=47 Hz, 27 Hz). Mass Spec (lo-res):Calc'd for C₇₇H₁₀₀FN₁₃O₂₃S₅: 1753.6. Found: 877.9 (M+2H⁺)/2.

Radiosynthesis of [18F]M005

A vial containing the click chemistry mixture of M005-precursor (2 mg),dissolved in 250 μL of 0.1 M CuSO₄, sodium ascorbate (20 mg), ACN (0.25mL) and EtOH/H₂O (2:1 ratio, 0.25 mL total). Aqueous ¹⁸F-fluoride ion,produced in the cyclotron target, was passed through an anion exchangeresin cartridge. The ¹⁸O—H₂O was retained for recycling. The ¹⁸Ffluoride was eluted from the column using a solution of potassiumcarbonate (3 mg) in water (0.4 mL) and collected into the reactionvessel. Kryptofix® 222 (20 mg) dissolved in acetonitrile (1 mL) wasadded to the aqueous ¹⁸F-fluoride in the reaction vessel. The mixturewas dried by heating between 68-95° C. under reduced pressure (250 mbar)and a stream of argon. After the ¹⁸F-fluoride was consideredsufficiently activated (as indicated by a constant pressure reading forthe reactor while under vacuum), the tosylate precursor was then addedas a solution in anhydrous MeCN (20 mg in 0.8 mL). The reaction washeated closed at 135° C. for 3-5 mM for fluorination. Afterfluorination, the transfer line in the reaction vessel was kept abovethe surface of the reaction mixture and opened for approximately 20seconds to allow the transfer of ¹⁸F-pentyne from the reaction pot intothe click vial (via bubbling of the labeled pentyne into the clicksolution) with the HPLC transfer line held above the reaction mixture.After the transfer was complete, the click reaction was allowed toproceed for 20 min. After clicking, the transfer line was lowered intothe click solution and the contents were pressurized through the HPLCload vial for dilution with water prior to purification. The reactionmixture containing crude ¹⁸F-M005 was transferred to the HPLC sampleloop (5 mL) and purified via chromatographic separation using asemi-preparative HPLC column (Phenomenex Gemini, C18, 5μ, 10×250 mm)using a isocredit 30% MeCN in 20 mM Phosphate buffer (pH=7.6) at 5mL/min. The column effluent was monitored using UV (210, 254 or 280 nm)and radiometric detectors connected in series. Purified ¹⁸F-M005 wascollected from the column at the retention time window determined forthe M005 reference standard, which coincides with the time that theradiometric detectors begin showing the main peak. The retention time ofthe ¹⁸F-M005 in this system was approximately 33 minutes. The purified¹⁸F-M005 fraction, eluted from the HPLC purification column, was dilutedwith water (40 mL) and captured onto a C18 SepPak cartridge. The C18SepPak cartridge was washed with water (10 mL) followed by elution ofthe product with 0.5 mL of EtOH. The sample was then diluted withsterile water (4.5 mL of water) to afford a final formulation of¹⁸F-M005 in a maximum of 10% EtOH:water. For sterile preparations, thedose was filtered through a sterile Pall 0.2 um filter.

Radiosynthesis of [18F]M005

Chemical syntheses of M005 precursor and reference standard.

In one embodiment, the invention is a compound of the Formula (I):

and pharmaceutically acceptable salts and stereoisomers thereof,wherein:

R₁ is selected from the group consisting of: C₁₋₆alkyl, C₁₋₆alkyl-halo,protecting group or leaving group;

A is a bond, (—CH₂—)₁₋₅, or is (—CH₂—CH₂—O—)₁₋₅;

R₃ is selected from the group consisting of: OH, NH₂, protecting groupor leaving group;

R₄ is a bond or is at least one selected from the group consisting of:aryl, heteroaryl,

-   -   C₁₋₂₀alkyl, wherein at least one C of C₁₋₂₀alkyl is optionally        replaced by O, S, NH, C(O), aryl or heteroaryl and wherein at        least one H of C₁₋₂₀alkyl is optionally replaced with OH, NH₂,        COOH, C₁₋₈alkyl, C₁₋₈alkyl-halo,        -   wherein at least one C of C₁₋₈alkyl is optionally replaced            by O, S, C(O), aryl, heteroaryl or NH,

R₅ is selected from the group consisting of: H, halo, CH₃, NH₂, OH, SO₂,NO₂, —O—C₁₋₆alkyl,

C₁₋₂₀alkyl, C₁₋₂₀alkyl-aryl, C₁₋₂₀alkyl-heteroaryl,

-   -   wherein at least one C of the C₁₋₂₀alkyl is optionally replaced        with S, SO₂, C(O), C(S), aryl, heteroaryl or NH,    -   wherein at least one H of the C₁₋₂₀alkyl, aryl or heteroaryl is        replaced with halo, OH, COOH, SO₃, NH₃, N(alkyl)₂, C₁₋₈alkyl,        C₁₋₈alkyl-halo,        -   wherein at least one C of C₁₋₈alkyl is optionally replaced            by O, S, C(O), aryl, heteroaryl or NH,

wherein at least one H or halo is optionally replaced with aradionuclide or a fluorophore.

In another embodiment, the invention is a compound of Formula (I)wherein A is a bond.

In another embodiment, the invention is a compound of Formula (I),wherein R₁ is C₁₋₆alkyl.

In another embodiment, the invention is a compound of Formula (I),wherein R₃ is OH.

In another embodiment, the invention is a compound of Formula (I),wherein R₅ is a halo or a radionuclide.

In another embodiment, the invention is a compound of Formula (I),wherein R₄ is C₁₋₂₀alkyl, wherein at least one C of C₁₋₂₀alkyl isoptionally replaced by O, S, NH, C(O).

In another embodiment, the invention is a compound of Formula (I), thatis:

In another embodiment, the invention is a compound of Formula (I), thatis:

In another embodiment, the invention is a compound of Formula (I), thatis:

wherein Z is a halo or a radionuclide.

In another embodiment, the invention is a compound of Formula (I), thatis:

wherein Z is a halo or a radionuclide.

In another embodiment, the invention is a compound of Formula (I),wherein R₄ is aryl.

In another embodiment, the invention is a compound of Formula (I),wherein R₄ is:

In another embodiment, the invention is a compound of Formula (I),wherein R₄ is C₁₋₂₀alkyl, wherein: at least one C of C₁₋₂₀alkyl isreplaced by 0, at least one C of C₁₋₂₀alkyl is replaced by C(O)NH, atleast one C of C₁₋₂₀alkyl is replaced by aryl, at least one C ofC₁₋₂₀alkyl is replaced by heteroaryl, wherein at least one H of the arylis replaced by COOH.

In another embodiment, the invention is a compound of Formula (I),wherein R₄ is —[(CH₂)₂—O]₁₋₆—.

In another embodiment, the invention is a compound of Formula (I),wherein R₅ is:

In another embodiment, the invention is a compound of Formula (I),wherein the heteroaryl is a triazole.

In another embodiment, the invention is a compound of Formula (I),wherein at least one H of C₁₋₂₀alkyl is replaced with

wherein Z is a halo or a radionuclide.

In another embodiment, the invention is a compound of Formula (I),wherein R₄ is

In another embodiment, the invention is a compound of Formula (II):

and pharmaceutically acceptable salts and stereoisomers thereof,wherein:each R₁ is independently selected from the group consisting of:C₁₋₆alkyl, C₁₋₆alkyl-halo, protecting group or leaving group;each A is a bond, (CH₂)₁₋₅, or is (—CH₂—CH₂—O—)₁₋₅;each R₃ is independently selected from the group consisting of: OH, NH₂,protecting group or leaving group;each R₆ is independently a bond or is at least one selected from thegroup consisting of: aryl, heteroaryl, orC₁₋₂₀alkyl, wherein at least one C of C₁₋₂₀alkyl is optionally replacedby O, S, NH, C(O), aryl or heteroaryl and wherein at least one H ofC₁₋₂₀alkyl is optionally replaced with OH, NH₂, C₁₋₈alkyl,C₁₋₈alkyl-halo,wherein at least one C of C₁₋₈alkyl is optionally replaced with at leastone of O, S, C(O), NH, aryl and heteroaryl;each R₇ is independently a bond or is at least one selected from thegroup consisting of: CH₂, NH, C(O)NH, C₁₋₆alkyl, wherein at least one Cof C₁₋₆alkyl is optionally replaced by O, S, NH, C(O) and wherein atleast one H of C₁₋₆alkyl is optionally replaced by OH, NH₂, SO₂,C₁₋₈alkyl, C₁₋₈alkyl-halo,wherein at least one C of C₁₋₈alkyl is optionally replaced by O, S,C(O), aryl, heteroaryl or NH,R₈ is a bond or is at least one selected from the group consisting of:O, NH, S, aryl, heteroaryl, wherein at least one H of aryl or heteroarylis optionally replaced with C₁₋₈alkyl, C₁₋₈alkyl halo, wherein at leastone C of C₁₋₈alkyl is optionally replaced with at least one of O, S,C(O), NH, aryl and heteroaryl;(CH₂)₁₋₆, wherein at least one C of (CH₂)₁₋₆ is optionally replaced byO, NH or S;wherein at least one H or halo is optionally replaced with aradionuclide or a fluorophore.

In another embodiment, the invention is a compound of Formula (II),wherein each A is a bond.

In another embodiment, the invention is a compound of Formula (II),wherein each R₃ is OH.

In another embodiment, the invention is a compound of Formula (II),wherein each R₁ is CH₃.

In another embodiment, the invention is a compound of Formula (II),wherein each R₆ is a bond.

In another embodiment, the invention is a compound of Formula (II),wherein R₈ is O.

In another embodiment, the invention is a compound of Formula (II),wherein each R₇ is C₁₋₆alkyl.

In another embodiment, the invention is a compound of Formula (II),wherein each R₆ is C₁₋₂₀alkyl, wherein at least one C of C₁₋₂₀alkyl isoptionally replaced by O, S, NH, C(O), aryl or heteroaryl and wherein atleast one H of C₁₋₂₀alkyl is optionally replaced with OH, NH₂,C₁₋₈alkyl-halo, wherein at least one C of C₁₋₈alkyl is optionallyreplaced with at least one of O, S, C(O), NH, aryl and heteroaryl.

In another embodiment, the invention is a compound of Formula (II),wherein each R₆ is —[(CH₂)₂—O]₁₋₆—.

In another embodiment, the invention is a compound of Formula (II),wherein each

In another embodiment, the invention is a compound of Formula (II),wherein each R₇ is C₁₋₆alkyl, wherein at least one C of C₁₋₆alkyl isoptionally replaced by O, S, NH, C(O) and at least one H is replacedwith:

wherein Z is a halo or a radionuclide.

In another embodiment, the invention is a compound of Formula (II),wherein each R₇ is CH₂C(O)NH.

In another embodiment, the invention is a compound of Formula (II),wherein R₈ is aryl.

In another embodiment, the invention is a compound of Formula (II),wherein at least one H of aryl is replaced with C₁₋₈alkyl-halo, whereinat least one C of C₁₋₈alkyl-halo is optionally replaced with heteroaryl.

In another embodiment, the invention is a compound of Formula (II), thatis:

and pharmaceutically acceptable salts and stereoisomers thereof.

In another embodiment, the invention is a compound of Formula (II) thatis:

and pharmaceutically acceptable salts and stereoisomers thereof,wherein Z is a halo or a radionuclide.

In another embodiment, the invention is a compound of Formula (II) thatis:

and pharmaceutically acceptable salts and stereoisomers thereof,wherein Z is a halo or a radionuclide.

In another embodiment, the invention is a compound of Formula (III):

and pharmaceutically acceptable salts and stereoisomers thereof,wherein:R₁ is selected from the group consisting of: C₁₋₆alkyl, C₁₋₆alkyl-halo,protecting group or leaving group;A is a bond, (CH₂)₁₋₅, or is (—CH₂—CH₂—O—)₁₋₅;R₃ is selected from the group consisting of: OH, NH₂, protecting groupor leaving group;

R₁₀ is selected from the group consisting of: an alkyne, an azide,C₁-C₆-alkyne, C₁-C₆-azide, wherein at least one C is optionally replacedby NH, C(O), S or O,

wherein at least one H or halo is optionally replaced with aradionuclide or a fluorophore.

In another embodiment, the invention is a compound of Formula (IV):

and pharmaceutically acceptable salts and stereoisomers thereof,wherein:R₁ is selected from the group consisting of: C₁₋₆alkyl, C₁₋₆alkyl-halo,protecting group or leaving group;A is a bond, (CH₂)₁₋₅, or is (—CH₂—CH₂—O—)₁₋₅;R₃ is selected from the group consisting of: OH, NH₂, protecting groupor leaving group;R₉ is a bond or is at least one selected from the group consisting of:aryl, heteroaryl,C₁₋₂₀alkyl, wherein at least one C of C₁₋₂₀alkyl is optionally replacedby O, S, NH, C(O), aryl or heteroaryl and wherein at least one H ofC₁₋₂₀alkyl is optionally replaced with OH, NH₂, COOH,wherein at least one C of C₁₋₈alkyl is optionally replaced by O, S,C(O), aryl, heteroaryl or NH,

R₁₀ is selected from the group consisting of: an alkyne, an azide,C₁-C₆-alkyne, C₁-C₆-azide, wherein at least one C is optionally replacedby NH, C(O), S or O,

wherein at least one H or halo is optionally replaced with aradionuclide or a fluorophore.

In another embodiment, the invention is a compound of Formula (V):

and pharmaceutically acceptable salts and stereoisomers thereof,wherein:R₁ is selected from the group consisting of: C₁₋₆alkyl, C₁₋₆alkyl-halo,protecting group or leaving group;A is a bond, (CH₂)₁₋₅, or is (—CH₂—CH₂—O—)₁₋₅;R₃ is selected from the group consisting of: OH, NH₂, protecting groupor leaving group;R₉ is a bond or is at least one selected from the group consisting of:aryl, heteroaryl,C₁₋₂₀alkyl, wherein at least one C of C₁₋₂₀alkyl is optionally replacedby O, S, NH, C(O), aryl or heteroaryl and wherein one H of C₁₋₂₀alkyl isoptionally replaced with OH, NH₂, COOH, C₁₋₈alkyl, C₁₋₈alkyl-halo,wherein at least one C of C₁₋₈alkyl is optionally replaced by O, S,C(O), aryl, heteroaryl or NH,

-   -   R₁₀ is selected from the group consisting of: an alkyne, an        azide, C₁-C₆-alkyne, C₁-C₆-azide, wherein at least one C of        C₁-C₆ is optionally replaced by NH, C(O), S or O;    -   R₁₁ is selected from the group consisting of: H, CH₃, NH₂, OH,        SO₂, NO₂, —O—C₁₋₆alkyl and

C₁₋₂₀alkyl, C₁₋₂₀alkyl-aryl, C₁₋₂₀alkyl-heteroaryl,wherein at least one C of the C₁₋₂₀alkyl is optionally replaced with S,SO₂, C(O), C(S), aryl, heteroaryl or NH,wherein at least one H of the C₁₋₂₀alkyl, aryl or heteroaryl is replacedwith halo, OH, COOH, SO₃, NH₃, N(alkyl)₂, C₁₋₈alkyl, C₁₋₈alkyl-halo,wherein at least one C of C₁₋₈alkyl is optionally replaced by O, S,C(O), aryl, heteroaryl or NH,wherein at least one H or halo is optionally replaced with aradionuclide or a fluorophore.

In another embodiment, the invention is a compound of Formula (VI):

and pharmaceutically acceptable salts and stereoisomers thereof,wherein:each R₁ is independently selected from the group consisting of:C₁₋₆alkyl, C₁₋₆alkyl-halo, protecting group or leaving group;each A is a bond, (CH₂)₁₋₅, or is (—CH₂—CH₂—O—)₁₋₅;each R₃ is independently selected from the group consisting of: OH, NH₂,protecting group or leaving group;each R₆ is independently a bond or is at least one selected from thegroup consisting of: aryl, heteroaryl, orC₁₋₂₀alkyl, wherein at least one C of C₁₋₂₀alkyl is optionally replacedby O, S, NH, C(O), aryl or heteroaryl and wherein at least one H ofC₁₋₂₀alkyl is optionally replaced with OH, NH₂, C₁₋₈alkyl,C₁₋₈alkyl-halo,wherein at least one C of C₁₋₈alkyl is optionally replaced with at leastone of O, S, C(O), NH, aryl and heteroaryl;each R₁₂ is independently a bond or is C₁₋₆alkyl, wherein at least one Cof C₁₋₆alkyl is optionally replaced by O, S, NH, C(O) and wherein atleast one H of C₁₋₆alkyl is optionally replaced by OH, NH₂, SO₂,C₁₋₈alkyl, C₁₋₈alkyl-halo, azide or alkyne,wherein at least one C of C₁₋₈alkyl is optionally replaced by O, S,C(O), aryl, heteroaryl or NH,R₈ is a bond or is at least one selected from the group consisting of:O, NH, S, aryl, heteroaryl, wherein at least one H of aryl or heteroarylis optionally replaced with C₁₋₈alkyl, C₁₋₈alkyl-halo,wherein at least one C of C₁₋₈alkyl is optionally replaced with at leastone of O, S, C(O), NH, aryl and heteroaryl;(CH₂)₁₋₆, wherein at least one C of (CH₂)₁₋₆ is optionally replaced byO, NH or S,wherein at least one H or halo is optionally replaced with aradionuclide or a fluorophore.

In another embodiment, the invention is a compound of Formula (VII):

and pharmaceutically acceptable salts and stereoisomers thereof,wherein:A is a bond, (CH₂)₁₋₅, or is (—CH₂—CH₂—O—)₁₋₅;R₃ is selected from the group consisting of: OH, NH₂, protecting groupor leaving group;R₄ is a bond or is at least one selected from the group consisting of:aryl, heteroaryl,C₁₋₂₀alkyl, wherein at least one C of C₁₋₂₀alkyl is optionally replacedby O, S, NH, C(O), aryl or heteroaryl and wherein at least one H ofC₁₋₂₀alkyl is optionally replaced with OH, NH₂, COOH, C₁₋₈alkyl,C₁₋₈alkyl-halo,wherein at least one C of C₁₋₈alkyl is optionally replaced by O, S,C(O), aryl, heteroaryl or NH,R₅ is selected from the group consisting of: H, halo, CH₃, NH₂, OH, SO₂,NO₂, —O—C₁₋₆alkyl and

C₁₋₂₀alkyl, C₁₋₂₀alkyl-aryl, C₁₋₂₀alkyl-heteroaryl,wherein at least one C of the C₁₋₂₀alkyl is optionally replaced with S,SO₂, C(O), C(S), aryl, heteroaryl or NH,wherein at least one H of the C₁₋₂₀alkyl, aryl or heteroaryl is replacedwith halo, OH, COOH, SO₃, NH₃, N(alkyl)₂, C₁₋₈alkyl, C₁₋₈alkyl-halo,wherein at least one C of C₁₋₈alkyl is optionally replaced by O, S,C(O), aryl, heteroaryl or NH,wherein at least one H or halo is optionally replaced with aradionuclide or a fluorophore.

In another embodiment, the invention is a compound of Formula (VIII):

and pharmaceutically acceptable salts and stereoisomers thereof,wherein:each A is a bond, (CH₂)₁₋₅, or is (—CH₂—CH₂—O—)₁₋₅;each R₃ is selected from the group consisting of: OH, NH₂, protectinggroup or leaving group;each R₄ is independently a bond or is at least one selected from thegroup consisting of: aryl, heteroaryl,C₁₋₂₀alkyl, wherein at least one C of C₁₋₂₀alkyl is optionally replacedby O, S, NH, C(O), aryl or heteroaryl and wherein at least one H ofC₁₋₂₀alkyl is optionally replaced with OH, NH₂, COOH, C₁₋₈alkyl,C₁₋₈alkyl-halo,wherein at least one C of C₁₋₈alkyl is optionally replaced by O, S,C(O), aryl, heteroaryl or NH,each R₅ is selected from the group consisting of: H, halo, CH₃, NH₂, OH,SO₂, NO₂, —O—C₁₋₆alkyl,

C₁₋₂₀alkyl, C₁₋₂₀alkyl-aryl, C₁₋₂₀alkyl-heteroaryl,wherein at least one C of the C₁₋₂₀alkyl is optionally replaced with S,SO₂, C(O), C(S), aryl, heteroaryl or NH,wherein at least one H of the C₁₋₂₀alkyl, aryl or heteroaryl is replacedwith halo, OH, COOH, SO₃, NH₃, N(alkyl)₂, C₁₋₈alkyl, C₁₋₈alkyl-halo,wherein at least one C of C₁₋₈alkyl is optionally replaced by O, S,C(O), aryl, heteroaryl or NH,R₁₃ is a bond or is C₁₋₂₀alkyl, wherein at least one C of the C₁₋₂₀alkylis optionally replaced with S, SO₂, C(O), C(S), aryl, heteroaryl or NH,wherein at least one H or halo is optionally replaced with aradionuclide or a fluorophore.

In another embodiment, the invention is a compound of Formula (IX):

and pharmaceutically acceptable salts and stereoisomers thereof,wherein:each R₁ is independently selected from the group consisting of:C₁₋₆alkyl, C₁₋₆alkyl-halo, protecting group or leaving group;A is a bond, (CH₂)₁₋₅, or is (—CH₂—CH₂—O—)₁₋₅;each R₃ is independently selected from the group consisting of: OH, NH₂,protecting group or leaving group;R₁₄ is independently a bond or is at least one selected from the groupconsisting of: aryl, heteroaryl, or C₁₋₈alkyl, wherein at least one C ofC₁₋₈alkyl is optionally replaced by O, S, NH, C(O), aryl or heteroaryl,wherein at least one H or halo is optionally replaced with aradionuclide or a fluorophore.

In another embodiment, the invention is a compound of Formula (X):

and pharmaceutically acceptable salts and stereoisomers thereof,wherein:R₁ is selected from the group consisting of: C₁₋₆alkyl, C₁₋₆alkyl-halo,protecting group or leaving group;A is a bond, (CH₂)₁₋₅, or is (—CH₂—CH₂—O—)₁₋₅;R₃ is selected from the group consisting of: OH, NH₂, protecting groupor leaving group;R₄ is a bond or is at least one selected from the group consisting of:aryl, heteroaryl,C₁₋₂₀alkyl, wherein at least one C of C₁₋₂₀alkyl is optionally replacedby O, S, NH, C(O), aryl or heteroaryl and wherein at least one H ofC₁₋₂₀alkyl is optionally replaced with OH, NH₂, COOH,wherein at least one C of C₁₋₈alkyl is optionally replaced by O, S,C(O), aryl, heteroaryl or NH,R₅ is selected from the group consisting of: H, halo, CH₃, NH₂, OH, SO₂,NO₂, —O—C₁₋₆alkyl,

C₁₋₂₀alkyl, C₁₋₂₀alkyl-aryl, C₁₋₂₀alkyl-heteroaryl,wherein at least one C of the C₁₋₂₀alkyl is optionally replaced with S,SO₂, C(O), C(S), aryl, heteroaryl or NH,wherein at least one H of the C₁₋₂₀alkyl, aryl or heteroaryl is replacedwith halo, OH, COOH, SO₃, NH₃, N(alkyl)₂, C₁₋₈alkyl, C₁₋₈alkyl-halo,wherein at least one C of C₁₋₈alkyl is optionally replaced by O, S,C(O), aryl, heteroaryl or NH, wherein at least one H or halo isoptionally replaced with a radionuclide or a fluorophore.

In another embodiment, the invention is a compound of Formula (X) thatis:

wherein Z is selected from the group consisting of: NH₂, OMe, halo orradionuclide.

While this invention has been described in conjunction with the specificembodiments outlined above, it is evident that many alternatives,modifications, and variations will be apparent to those skilled in theart. Accordingly, the preferred embodiments of the invention as setforth above are intended to be illustrative, not limiting. A variety ofmodifications to the embodiments described will be apparent to thoseskilled in the art from the disclosure provided herein. Thus, thepresent invention may be embodied in other specific forms withoutdeparting from the spirit or essential attributes thereof. Variouschanges may be made without departing from the spirit and scope of theinventions as defined in the following claims.

We claim:
 1. A compound of Formula (I):

and pharmaceutically acceptable salts and stereoisomers thereof,wherein: R₁ is selected from the group consisting of: C₁₋₆alkyl,C₁₋₆alkyl-halo, protecting group or leaving group; A is a bond,(—CH₂—)₁₋₅, or is (—CH₂—CH₂—O—)₁₋₅; R₃ is selected from the groupconsisting of: OH, NH₂, protecting group or leaving group; R₄ is a bondor is at least one selected from the group consisting of: aryl,heteroaryl, C₁₋₂₀alkyl, wherein at least one C of C₁₋₂₀alkyl isoptionally replaced by O, S, NH, C(O), aryl or heteroaryl and wherein atleast one H of C₁₋₂₀alkyl is optionally replaced with OH, NH₂, COOH,wherein at least one C of C₁₋₈alkyl is optionally replaced by O, S,C(O), aryl, heteroaryl or NH, R₅ is selected from the group consistingof: H, halo, CH₃, NH₂, OH, SO₂, NO₂, —O—C₁₋₆alkyl,

C₁₋₂₀alkyl, C₁₋₂₀alkyl-aryl, C₁₋₂₀alkyl-heteroaryl, wherein at least oneC of the C₁₋₂₀alkyl is optionally replaced with S, SO₂, C(O), C(S),aryl, heteroaryl or NH, wherein at least one H of the C₁₋₂₀alkyl, arylor heteroaryl is replaced with halo, OH, COOH, SO₃, NH₃, N(alkyl)₂,C₁₋₈alkyl, C₁₋₈alkyl-halo, wherein at least one C of C₁₋₈alkyl isoptionally replaced by O, S, C(O), aryl, heteroaryl or NH, wherein atleast one H or halo is optionally replaced with a radionuclide or afluorophore.
 2. The compound of claim 1, wherein A is a bond.
 3. Thecompound of claim 1, wherein R₁ is C₁₋₆alkyl.
 4. The compound of claim1, wherein R₃ is OH.
 5. The compound of claim 1, wherein R₅ is a halo ora radionuclide.
 6. The compound of claim 1, wherein R₄ is C₁₋₂₀alkyl,wherein at least one C of C₁₋₂₀alkyl is optionally replaced by O, S, NH,C(O).
 7. The compound of claim 1 that is:


8. The compound of claim 1 that is:


9. The compound of claim 1 that is:

wherein Z is a halo or a radionuclide.
 10. The compound of claim 1 thatis:

wherein Z is a halo or a radionuclide.
 11. The compound of claim 5,wherein R₄ is aryl.
 12. The compound of claim 5, wherein R₄ is:


13. The compound of claim 5, wherein R₄ is C₁₋₂₀alkyl, wherein: at leastone C of C₁₋₂₀alkyl is replaced by 0, at least one C of C₁₋₂₀alkyl isreplaced by C(O)NH, at least one C of C₁₋₂₀alkyl is replaced by aryl, atleast one C of C₁₋₂₀alkyl is replaced by heteroaryl, wherein at leastone H of the aryl is replaced by COOH.
 14. The compound of 6, wherein R₄is —[(CH₂)₂—O]₁₋₆—.
 15. The compound of claim 6, wherein R₅ is:


16. The compound of claim 13, wherein the heteroaryl is a triazole. 17.The compound of claim 15, wherein at least one H of C₁₋₂₀alkyl isreplaced with

wherein Z is a halo or a radionuclide.
 18. The compound of claim 17,wherein R₄ is


19. A compound of Formula (V):

and pharmaceutically acceptable salts and stereoisomers thereof,wherein: R₁ is selected from the group consisting of: C₁₋₆alkyl,C₁₋₆alkyl-halo, protecting group or leaving group; A is a bond,(CH₂)₁₋₅, or is (—CH₂—CH₂—O—)₁₋₅; R₃ is selected from the groupconsisting of: OH, NH₂, protecting group or leaving group; R₉ is a bondor is at least one selected from the group consisting of: aryl,heteroaryl, C₁₋₂₀alkyl, wherein at least one C of C₁₋₂₀alkyl isoptionally replaced by O, S, NH, C(O), aryl or heteroaryl and whereinone H of C₁₋₂₀alkyl is optionally replaced with OH, NH₂, COOH,C₁₋₈alkyl, C₁₋₈alkyl-halo, wherein at least one C of C₁₋₈alkyl isoptionally replaced by O, S, C(O), aryl, heteroaryl or NH, R₁₀ isselected from the group consisting of: an alkyne, an azide,C₁-C₆-alkyne, C₁-C₆-azide, wherein at least one C of C₁-C₆ is optionallyreplaced by NH, C(O), S or O; R₁₁ is selected from the group consistingof: H, CH₃, NH₂, OH, SO₂, NO₂, —O—C₁₋₆alkyl and

C₁₋₂₀alkyl, C₁₋₂₀alkyl-aryl, C₁₋₂₀alkyl-heteroaryl, wherein at least oneC of the C₁₋₂₀alkyl is optionally replaced with S, SO₂, C(O), C(S),aryl, heteroaryl or NH, wherein at least one H of the C₁₋₂₀alkyl, arylor heteroaryl is replaced with halo, OH, COOH, SO₃, NH₃, N(alkyl)₂,C₁₋₈alkyl, C₁₋₈alkyl-halo, wherein at least one C of C₁₋₈alkyl isoptionally replaced by O, S, C(O), aryl, heteroaryl or NH, wherein atleast one H or halo is optionally replaced with a radionuclide or afluorophore.
 20. The compound of claim 19, that is:


21. A compound of Formula (IA):

wherein X₁ is selected from the group consisting of halo, fluorophore orradioactive halo.
 22. The compound of claim 21, wherein X₁ is F.
 23. Thecompound of claim 21, wherein X₁ is ¹⁸F.